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Genome-Scale CRISPR/Cas9 Screening Reveals Squalene Epoxidase as a Susceptibility Factor for Cytotoxicity of

Yukio Koizumi1, Jun Fukushima2, Yayoi Kobayashi2

  • 1Department of Biochemistry and Metabolic Science, Akita University Graduate School of Medicine, 1-1-1 Hondo, Akita 010-8543, Japan.

Chembiochem : a European Journal of Chemical Biology
|February 9, 2019
PubMed

Insights

Malformin A1 (MA1) is cytotoxic to mammalian cells. Researchers identified squalene epoxidase as a key factor in MA1

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Pharmacology

Background:

  • Malformin A1 (MA1), a cyclic pentapeptide from fungi, demonstrates cytotoxicity to mammalian cells.
  • Understanding the cytotoxic mechanism of MA1 is crucial for potential therapeutic applications.

Purpose of the Study:

  • To elucidate the molecular mechanism underlying Malformin A1-induced cytotoxicity.
  • To identify host genes mediating sensitivity or resistance to MA1.

Main Methods:

  • Utilized a CRISPR/Cas9 genome-wide knockout library for screening in U937 cells.
  • Employed positive selection for MA1-resistant cells.
  • Analyzed enriched single guide RNAs (sgRNAs) via high-throughput sequencing.

Main Results:

  • Squalene epoxidase (SQLE) was identified as a critical gene involved in MA1 cytotoxicity.
  • Depletion of SQLE conferred resistance to MA1 in U937 cells.
  • Inhibition of squalene epoxidase also rendered wild-type cells resistant to MA1.

Conclusions:

  • Squalene epoxidase plays a significant role in the cytotoxic effects of Malformin A1.
  • This discovery offers novel insights into potential therapeutic strategies for ischemic diseases using MA1.
Keywords:
CRISPR/Cas9cytotoxicitymalforminpeptidessqualene epoxidase

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